NCI-H295R [H295R] (ATCC® CRL-2128)

Organism: Homo sapiens, human  /  Tissue: adrenal gland/cortex  /  Disease: Carcinoma

Organism Homo sapiens, human
Tissue adrenal gland/cortex
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease Carcinoma
Age 48 years
Gender female
Ethnicity Black
Applications
This line has the ability to produce adrenal androgens.
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
NCI-H295R was adapted from the NCI-H295 pluripotent adrenocortical carcinoma cell line (ATCC CRL-10296) established by A.F. Gazdar and associates from a carcinoma of the adrenal cortex. The original cells were adapted to a culture medium which decreased the population doubling time from 5 days to 2 days. While the original cells grew in suspension, the adapted cells were selected to grow in a monolayer. This cell line retains the ability to produce adrenal androgens. It is responsive to angiotensin II and potassium ions.
Clinical Data
48 years
Black
female
Genes Expressed
aldosterone; cortisol; C19 steroids
Cellular Products
aldosterone; cortisol; C19 steroids
Comments
It is responsive to angiotensin II and potassium ions.
This cell line retains the ability to produce adrenal androgens.
Complete Growth Medium The base medium for this cell line is ATCC-formulated DMEM:F12 Medium Catalog No. 30-2006. To make the complete growth medium, add the following components to the base medium: 0.00625 mg/ml insulin; 0.00625 mg/ml transferrin; 6.25 ng/ml selenium; 1.25 mg/ml bovine serum albumin; 0.00535 mg/ml linoleic acid; adjust to a final concentration of 2.5% Nu-Serum I. The additives (insulin, transferrin, selenium, BSA and linoleic acid) are available in the form of ITS+ Premix from BD Biosciences. Nu-Serum is also available from BD Biosciences.
Subculturing Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
  6. Resuspend the cell pellet in fresh growth medium.  Add appropriate aliquots of cell suspension to new culture vessels.
  7. Incubate cultures at 37°C.

Subculture Ratio: 1:3 to 1:4
Medium Renewal: 2 to 3 times a week.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 4th edition, published by Wiley-Liss, N.Y., 2004.

Cryopreservation
Freeze medium: Complete growth medium + extra 7.5% Nu-Serum, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
STR Profile
Amelogenin: X
CSF1PO: 10,12
D13S317: 13
D16S539: 11
D5S818: 12
D7S820: 9,12
THO1: 9.3
TPOX: 8
vWA: 17,18
Name of Depositor WE Rainey
References

Rainey WE, et al. The NCI-H295 cell line: a pluripotent model for human adrenocortical studies. Mol. Cell. Endocrinol. 100: 45-50, 1994. PubMed: 8056157

Rainey WE, et al. Regulation of human adrenal carcinoma cell (NCI-H295) production of C19 steroids. J. Clin. Endocrinol. Metab. 77: 731-737, 1993. PubMed: 8396576

Gazdar AF, et al. Establishment and characterization of a human adrenocortical carcinoma cell line that expresses multiple pathways of steroid biosynthesis. Cancer Res. 50: 5488-5496, 1990. PubMed: 2386954

Bird IM, et al. Human NCI-H295 adrenocortical carcinoma cells: a model for angiotensin- II-responsive aldosterone secretion. Endocrinology 133: 1555-1561, 1993. PubMed: 8404594

Basic Documentation
Other Documentation
References

Rainey WE, et al. The NCI-H295 cell line: a pluripotent model for human adrenocortical studies. Mol. Cell. Endocrinol. 100: 45-50, 1994. PubMed: 8056157

Rainey WE, et al. Regulation of human adrenal carcinoma cell (NCI-H295) production of C19 steroids. J. Clin. Endocrinol. Metab. 77: 731-737, 1993. PubMed: 8396576

Gazdar AF, et al. Establishment and characterization of a human adrenocortical carcinoma cell line that expresses multiple pathways of steroid biosynthesis. Cancer Res. 50: 5488-5496, 1990. PubMed: 2386954

Bird IM, et al. Human NCI-H295 adrenocortical carcinoma cells: a model for angiotensin- II-responsive aldosterone secretion. Endocrinology 133: 1555-1561, 1993. PubMed: 8404594